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Description
Human MIP4a ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Macrophage Inflammatory Protein 4 Alpha (MIP4a). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Macrophage Inflammatory Protein 4 Alpha (MIP4a) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Macrophage Inflammatory Protein 4 Alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Macrophage inflammatory protein 4α (MIP4a) is a small cytokine belonging to the CC chemokine family. It is also known as Eotaxin-3, chemokine (CC pattern) ligand 26, thymic stromal chemokine 1 (TSC-1), and IMAC. It is expressed in several tissues, including the heart, lung, and ovary, as well as in endothelial cells stimulated by the cytokine interleukin-4. Diseases associated with CCL26 include esophagitis and Churg-Strauss syndrome. Pathways involved include the TGF-β pathway and PEDF-induced signaling. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
Good product
Flavor Name: Spearmint, Size: 55 Count (Pack of 1)
Very soft and chewy, sugar free and has good flavor. It does lose its flavor around 10 minutes in but that's how long its meant to be chewed for anyway.
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Reviewed in the United States on May 16, 2026
★★★★★ 5
Great for your teeth and breath
Flavor Name: Spearmint, Size: 55 Count (Pack of 1)
Purchased this brand because it claimed to have no aspartame and was 100% xylitol. Xylitol is claimed to be a teeth remineralizer and I want fresh breath without all of the sugar. These are a nice, mild mint. Some mint gums make your eyes water they are so strong, this is definitely a nice range for me- did not make my eyes water but does give you nice breath. Flavor lasts a long time. I like it and I like that the pouch is resealable for carrying or the little chiclet style pieces can be placed in an aesthetic tin in your bag for on the go.
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Reviewed in the United States on May 2, 2026
★★★★★ 4
Perfect Keto-Friendly Gum, Clean Ingredients & Great Flavor!
Flavor Name: Spearmint, Size: 55 Count (Pack of 1), Flavor Name: Spearmint, Size: 55 Count (Pack of 1)
I’ve tried so many sugar-free gums, and this is by far the best option for anyone following a keto or strict carnivore lifestyle. It’s aspartame-free, has clean ingredients, and doesn’t leave that artificial aftertaste that most gums have. The spearmint flavor is fresh and long-lasting, and I love that it helps satisfy cravings without adding sugar or unnecessary carbs.
The texture stays soft longer than other brands, and the resealable bag is super convenient for keeping in the car, gym bag, or at work. If you’re looking for a healthier gum that fits perfectly into a low-carb, keto, or carnivore-focused routine, this is definitely the one to buy. Four stars only because the flavor only lasts about a minute.
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Reviewed in the United States on May 21, 2026
★★★★★ 5
Good alternative to traditional gums- no aspartame no artificial coloring.
Flavor Name: Spearmint, Size: 55 Count (Pack of 1)
Love this gum, no aspartame, no artificial coloring and good taste, you get used to chewing it for a long time, the taste goes away little faster than traditional gums but I don't care
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Reviewed in the United States on May 28, 2026
★★★★★ 5
Great Xylitol Chewing Gum!
Flavor Name: Spearmint, Size: 55 Count (Pack of 3), Flavor Name: Spearmint, Size: 55 Count (Pack of 3)
This review is for PUR Gum | Aspartame Free Chewing Gum | 100% Xylitol | Natural Spearmint Flavored Gum, 55 Pieces (Pack of 3). I got 2 of the 3-pack at the time because it was cheaper to do so than to get a 6-pack. Of course, the next day the 6-pack was cheaper lol. Anyway, this is great xylitol gum and not overly expensive like Zellie’s. I take 2 (which is one serving meaning 2g of xylitol) after “most” meals because it’s supposed to be good for my oral health. I chew one piece at a time for a couple minutes to prolong the sweet flavor instead of popping both into my mouth together. I like chewing these because they don’t harden up like some gums do. They stay soft throughout and I often forget and chew them for 15-20 minutes at a time. Anyway, I did the math and compared the PUR gum to other 100% xylitol gums out there and this one was a clear winner to me for the same benefits. I would totally recommend this gum and I like that it’s aspartame free! I’d give it a 5.0/5.0 - 03/20/2026.
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Reviewed in the United States on March 20, 2026